Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P30536 (PBS)
 9,886 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Neutrophils from bovine milk and blood platelets from dog plasma were washed in PBS, fixed in GA, dehydrated, suspended in a drop on a formvar-coated slide and immediately critical-point-dried in CO2. After coating with Pt-Pd the specimens were examined in an SEM. The same cells were then examined by interferometry (Int) in a light microscope, and the dry mass was determined. It is shown that this preparation method for both types of microscopes (SEM and Int) appears to give adequate results as far as fine surface structure (SEM-appearance) and dry mass determinations (Int) are concerned. The method has the advantage of a more precise characterization of individual particles, than would have been possible, if both methods of microscopy (SEM and Int) had been employed on the same sample, but on different specimens.
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PMID:Preparation of cells from suspensions for correlative scanning electron and interference microscopy. 110 40

Human follicular fluid collected by laparoscopic oocyte pick-up during IVF was studied with a computer-assisted semen analyser to evaluate the effect of hFF on human sperm motility and velocity. Freshly ejaculated human sperm were washed with phosphate buffered saline and mixed with either PBS or hFF. At various incubation periods of time, hFF increased both sperm motility and velocity as compared with control (P less than 0.01). After incubation of sperm with hFF at 37 degrees C and 5% CO2 in air for 0, 1, 3, 6, and 12 h, the amplitude increase of motility were 49%, 77%, 330%, 2020%, and 3340% when individual control motility was considered to be 100%. The amplitude increase of curvilinear velocity were 43%, 51%, 67%, 152%, 278%, respectively. Comparison of the motility and velocity of the sperm treated with hFF between 0 and 12 h, showed that hFF preserved both motility and velocity in vitro (P less than 0.01). The stimulatory effect of hFF was retained after boiling at 100 degrees C for 30 min, or after being filtered through Amicon membrane cones, but it disappeared if the hFF had been pre-treated with chymotrypsin. However, hFF did not stimulate the motility and velocity of unwashed sperm in freshly ejaculated human semen. A non-dialyzable and heat-stable factor(s) with a molecular weight below 50,000 in hFF may improve and maintain the motility and velocity of washed human sperm. Whether this factor could be used to improve pregnancy rate in assisted reproduction awaits further investigation.
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PMID:Human follicular fluid stimulates motility and velocity of washed human sperm in vitro. 151 76

The investigations were carried out on 59 cows from Holstein half-breed, establishing that 8 cows suffered salpinx obstruction (5 cases with unilateral obstruction and 3 cases with bilateral obstruction). The authors are using an apparatus made by themselves, for insufflation of air in the obstructed uterus, and which is useful in desobstruction treatment, too. For diagnosis, CO2 was introduced inside of uterus. The authors used for treatment PBS (saline phosphate buffer) in addition with penicillin G, hydrocortisone and trypsin. Before air insufflation in uterus there will be infused 10-20 ml 2% Lidocaton. The cows must be examined in oestrus period, or 2 days after PGF2 alfa administration. The gas must be introduced under rectal palpation, and pressure must not be higher than 500 mm H2O column. If there is a permeable oviduct, after 15-20 sec. from gas introduction, ist is possible to palpate the filled oviduct. From ovary we can perceive a rustle produced by gas crossing in abdominal cavity. In case of salpinx obstruction, the treatment must be start as soon as possible. The utilized liquid for treatment will be introduce by gas pressure, inside of uterus and oviducts. Using this method, it managed the repermeability of oviducts at 3 from 8 treated cows. In each case, there were used 3 treatments at 48 h interval. After the second insemination (I.A.) 2 cows remained pregnant.
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PMID:[A method for the treatment of oviduct obstruction in cattle using CO2 insufflation into the uterus]. 175 12

The aim of this study was to develop non-radioactive cell line proliferation assays. The human leukemic cell line TF1 (Kitamura et al., 1989) was used for the determination of the specific biological activity of recombinant human (rhu) granulocyte-macrophage colony-stimulating factor (GM-CSF) and rhu Interleukin 3 (IL-3) by a simple and economical fluorometric assay with a sensitivity similar to the measurement of 3H-thymidine uptake. The TF1 cell line responds to rhu IL-3, rhu GM-CSF and to a lesser extent to rhu Erythropoietin (EPO) and mast cell growth factor (MGF), but not to rhu G-CSF. It is dependent upon rhu GM-CSF for survival in culture. For the proliferation assay 1 x 10(4) TF1 cells were incubated with 20 ng - 0.256 pg rhu GM-CSF or rhu IL-3 at 37 degrees C and 5% CO2 in humidified atmosphere. After 48 h the cells were washed twice with PBS and were incubated with 4-Methylumbelliferyl-heptanoate for 60 min. Fluorescence was determined on a Titertek Fluoroskan II (Flow Lab.), and results were given as fluorescence units using a 355 nm excitation filter and a 480 nm emission filter. The developed assay showed an interassay variability lower than 15%. The sensitivity of the proliferation assays in the same range as the thymidine incorporation assays.
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PMID:Development of a rapid, highly sensitive, non-radioactive assay system for hematopoietic growth factors. 180 97

Four cell embryos collected by laparatomy from Sardinian breed ewes superovulated with FSH-p (16 mg Sigma), were divested of their zonae pellucidae (ZP) by micromanipulation or chemical methods (pronase 0.5%, tyrode pH 2.2). The blastomeres were separated by pipetting using a flame polished pasteur pipette in a Ca free medium (PBS. Sigma) and were inserted into previously evacuated Z.P. using a Leitz micromanipulator. The Z.P. were removed either mechanically or with acid tyrode; pronase was unable to digest them after incubation at 30 degrees C for 120 minutes. The single blastomeres were cocultured on a monolayer of ovine oviductal epithelial cells in TCM 199 + 10 FCS at 38 degrees C in 5% CO2 for 60 hours. No developments were observed in blastomeres obtained by acid digestion of the ZP while 50% of the other blastomeres continued their development until the 16 cell stages. Our results suggest that coculture with oviductal epithelial cell monolayers can support in vitro development of single ovine blastomeres.
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PMID:[In vitro separation and development of sheep blastomeres]. 210 May 29

6 days old bovine embryos (n = 126) were obtained from 8 superovulated cows or heifers by flushing the uteri and oviducts either non-surgically or after slaughter. Part of the embryos (n = 72) (morula stages) were placed in Ham's F-10 or PBS supplemented with 10% fetal calf serum (FCS) diluted 1:1 with supernatant from the H-Y antibody producing clone and cultured at 38 degrees C, in 5% CO2/95% air and 100% humidity. Control embryos (n = 54) were cultured in H-Y antibody free medium. After culture the embryos could be separated into a blastocyst--and a morula group. A subsequent colchemid and hypotonic treatment and fixation and Giemsa staining allowed a precise karyotyping, and thus sex determination for 36 H-Y antibody treated embryos and 22 control embryos. The limiting factor for proper karyotyping was lack of metaphases, incomplete methaphases or poor preparation. Among the H-Y antibody treated embryos we found 7 males and 15 females in the blastocyst and 14 males and 0 females in the morula group. A statistical analysis of these proportions led to the conclusion that the H-Y antibody had a significant influence on the sex ratio.
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PMID:Sex determination of bovine embryos using H-Y antibodies. 259 83

Embryos (8-16 cell) were obtained from random bred albino mice (6-8 weeks old) that were induced to superovulate by injections of 5 I.U. PMSG and 5 I.U. hCG given 48 hr apart. Embryos were exposed to intracellular cryoprotecting medium (glycerol 10%, 1-2 propanediol 20% in PBS) for 10 min and then transferred to extracellular vitrification medium (25% glycerol, 25% 1-2 propanediol in PBS). Vitrification medium containing embryos, and diluent (1 M sucrose) were loaded in a straw and immediately plunged into liquid N2. After thawing at 20 degrees C, the contents of the straw were mixed by shaking (1 step dilution) and emptied in a petri dish. After 3 washings in culture medium the embryos were kept in CO2 incubator for further development. In 3-step dilution procedure the dilution of cryoprotectants was done in 0.5 and 0.25 M sucrose before culture. Embryos in 3-step dilution of cryoprotectants exhibited high survival as compared to 1-step dilution (20.23% vs 6.55%).
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PMID:Cryopreservation of mouse embryos at -196 degrees C by vitrification. 280 16

Monolayer cultures of human thyroid cells derived from thyroid adenoma were utilized for the assay of thyroid stimulating substances such as thyrotropin (TSH), cholera toxin and thyroid stimulating immunoglobulin (TSI) in patients with Graves' disease. Adenoma cells were treated with 0.1% collagenase or 2000 unit/ml dispase to thyrocytes. The cells were cultured in MEM containing 10% fetal calf serum under an atmosphere of 5% CO2 in air. Within 24 hours, the cells attached themselves to the plastic surface and formed a monolayer. Cyclic AMP responses to TSH, cholera toxin or Graves' IgG were tested in a medium (PBS) containing 0.5 mM IBMX. The cyclic AMP responses to TSH were generally maximal on the 3rd day of culture and declined thereafter. The response was dose-dependent, and 10 microU/ml of TSH produced a significant increase of cellular cyclic AMP. The response by 1 microU/ml of TSH was 28 approximately 57 fold above the basal. The response was also a function of the incubation period. The maximal response was attained after 1 h incubation. When the cultures were washed after exposure to TSH, the cellular cyclic AMP levels rapidly declined, suggesting that removal of receptor-bound TSH results in a prompt cessation of cyclic AMP production. The thyroid cells in monolayer also responded to cholera toxin. The response was dose-dependent, and cholera toxin as low as 1 ng/ml was able to increase cyclic AMP production. In contrast to the observations in TSH, the cyclic AMP responses induced by cholera were hardly affected by washing the cultures after exposure to cholera toxin. Treatment of the cells with cholera toxin for only 3 min resulted in a continuous stimulation of cyclic AMP production for more than 4 hours. Confirming recent observations by others, most of Graves' IgG stimulated cyclic AMP production in a dose-dependent manner, but some of them inhibited the response at high concentrations. IgG derived from normal subjects did not increase cellular cyclic AMP. The time course in the cyclic AMP responses induced by Graves' IgG was variable among the IgG preparations from different patients. In some patients, the maximal responses were attained after 4 hours of incubation. A significant difference was noted between TSH and Graves' IgG in the stimulation of cyclic AMP production after washing the cultures. When the cultures were treated with Graves' IgG for 30 min, washed and then incubated without Graves' IgG, cellular cyclic AMP levels remained at the levels which were almost equivalent to those observed in the continuous presence of the IgGs.(ABSTRACT TRUNCATED AT 400 WORDS)
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PMID:[The effects of TSH, cholera toxin and Graves' IgG on cAMP production in cultured human thyroid adenoma cells in monolayer]. 286 66

In 2 radioimmunoassays in use to detect antibodies to dsDNA, the Farr assay and the PEG assay, we observed inhibitory effects of normal human serum (NHS) on the DNA binding by SLE sera. This was found to be due by the fact that, during incubation at 37 degrees C, CO2, introduced in the incubation mixture by the serum, evaporates from the mixture. This results in increase in pH to values well above pH 8.0, which in turn leads to a decreased DNA binding by antibody. When SLE sera are tested at low dilution, this phenomenon may lead to false negative results. Proper pH control, by the use of buffers with a greater buffering capacity than PBS, completely prevented the observed inhibitory effects. However, under these conditions NHS bound significant amounts of DNA in both assays. The non-specific DNA binding by NHS was found to be heat-stable, but could be eliminated either by aerosil treatment of the sera or by addition of dextran sulphate to the incubation mixture. Lipoproteins and, to a lesser extent, the complement component C1q appear responsible for this non-specific binding. To avoid false negative results with SLE sera as well as non-specific binding by NHS, we propose the use of stronger buffers in combination with added dextran sulphate to the incubation mixture in both the Farr assay and the PEG assay.
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PMID:Influence of pH on the detection of low- and high-avidity anti-dsDNA. 618 61

Cytotoxicity of an antiseptic is usually evaluated by microscopic examination of a cell culture, after a set time of contact with the antiseptic. As this evaluation is largely subjective, a photometric method is proposed. The procedure consists in staining the cells by methylene blue after contact with the ATS and measuring the dye in a photometer after elution. Different dilutions of ATS are added to a 24 h-monolayer of Vero cells in a 96-well microtissue culture plate (8 wells for each dilution, 8 wells for cell control, and 8 wells for control of dye fixation by the plastic plate). The plate is incubated for three days at 37 degrees C in an atmosphere of 5% CO2 and 95% air. The plate is washed with PBS to remove dead cells and adherent cells are fixed. The plate is then washed with borate buffer and allowed to dry. The dye is eluted by adding 200 microliter of HCl 0.1 N to each well. The plate is then read automatically at 650 nm on a Titertek Multiskan, a vertical light path photometer. Cytotoxicity is expressed as the percentage of damaged cells as compared to control wells. Cytotoxic assay of glutaraldehyde shows that this technique is more reliable and more sensitive than microscopic examination. Moreover, result of cytotoxic assay and of a method consisting in measurement of protein in residual cells after exposure to ATS are significantly correlated.
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PMID:[Evaluation of the cytotoxicity of an antiseptic by a photometric micromethod]. 643 83


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